Molecular cloning and expression of kidney‐type glutaminase from common carp (Cyprinus carpio) and its up‐regulation by glutamine in primary culture enterocyte

Glutaminase (GLS) is the key enzyme of glutamine (Gln) metabolism and utilization. In this study, a cDNA encoding GLS protein was identified from common carp Cyprinus carpio intestine. The open reading frame of GLS cDNA encodes a polypeptide of 595 amino acids, which shows a high similarity with its zebrafish Danio rerio counterpart. Bioinformatic analysis showed the protein belongs to kidney‐type GLS. The putative protein has glutaminase domain and ankyrin repeats domain, which are highly conserved among vertebrate orthologues. Real‐time quantitative PCR analysis revealed that the abundance of GLS mRNA was the highest in the white muscle, followed by the brain, eyeball and pituitary. Glutaminase was ubiquitously expressed in all intestinal segments of common carp. The activity of GLS did not distribute uniformly along the entire length of the intestine. In primary culture enterocyte, and the expression of GLS mRNA is up‐regulated quickly and effectively by Gln.     

Genetic variability within Phaeoisariopsis griseola from Central America and its implications for resistance breeding of common bean

The genetic and virulence variability of 112 isolates of Phaeoisariopsis griseola , collected from various locations in Central America, were studied using seven random amplified polymorphic DNA (RAPD) primers and 12 common-bean differential genotypes. Broad molecular diversity ( H  = 0·92) among isolates was found using RAPD markers. Fifty pathotypes were identified on 12 differential bean genotypes, 29 of which were represented by only one isolate. Only 18 pathotypes were found in two or more countries. Pathotype 63-63 was the most virulent and caused leaf spots on all 12 common-bean differential genotypes. Comparison of virulence phenotypes and RAPD profiles to known Andean P. griseola isolates confirmed that all isolates belonged to the Mesoamerican group. Pairwise comparison between individual RAPD loci showed that the majority were in gametic phase linkage disequilibrium, revealing that P. griseola maintains a genetic structure that is consistent with asexual reproduction. The molecular and virulence diversities of P. griseola isolates from Central America imply that using single resistance genes to manage angular leaf spot is inadequate and stacking resistance genes may be necessary to manage the disease effectively.     

Bacterial community composition of sediments from a milkfish Chanos chanos Forsskål farm

Bacterial communities in sediments underneath milkfish cages and adjacent off‐cage areas in Bolinao, Lingayen Gulf, West Philippine Sea were characterized using PCR‐denaturing gradient gel electrophoresis to assess influence of the fish farming to the sediments. The sediments in the fish cage (FC) area showed anoxic conditions [redox potential of ?165 to ?213 mV and acid‐volatile sulphide‐sulphur (AVS‐S) of 0.55–1.88 mg g^?1 (dry weight)] compared to the relatively oxic off‐cage area [redox potential of 72–81 mV and AVS‐S of 0.09–0.14 mg g^?1 (dry weight)]. The composition of the microbial populations revealed influence of the fish feed sedimentation with higher dominance of putative sulphate‐reducing bacteria (SRB) in the FC area. Compositions of the bacterial taxa showed dominance of the Bacteroidetes group and Deltaproteobacteria. The results suggest that the fish feeds created a new suitable environment, in which reductive microorganisms such as SRB, ferric‐ion‐reducing bacteria and nitrate‐reducing bacteria can thrive. Organic load from the feed and quantity of its leftover constituents may be the key factors that influence the structure of the bacterial community in the sediments of the milkfish farms.     

Effects of different dietary protein and lipid levels on growth, feed utilization and body composition of red porgy (Pagrus pagrus) fingerlings

Two feeding trials were conducted to determine the minimum dietary protein level producing maximum growth, and the optimum protein to energy ratio in diets for red porgy (Pagrus pagrus) fingerlings, respectively. In the first trial, six isoenergetic diets were formulated with protein levels ranging from 400 to 650 g kg^?1 in increments of 50 g kg^?1, and fed for 11 weeks to 2.8 g average initial weight fish. Weight gain, specific growth rate and feed efficiency were significantly higher with diets containing higher protein levels, when compared with dietary levels below 500 g kg^?1. The highest protein efficiency ratios were obtained in fish fed 500 g kg^?1 dietary protein. The minimum dietary protein level producing maximum fish growth was found to be 500 g kg^?1. In the second trial, 15 g average initial weight fish were fed for 12 weeks, six diets containing three different lipid levels (100, 150 and 200 g kg^?1) combined with two protein levels (450 and 500 g kg^?1). Weight gain values increased when dietary lipids increased from 100 to 150 g kg^?1, with a further decrease for 200 g kg^?1 lipids in diets; the lowest fish growth being supported by 200 g kg^?1 dietary lipids. Fish growth was significantly higher when dietary protein increased from 450 to 500 g kg^?1. There was no evidence of a protein‐sparing effect of dietary lipids. Liver protein and lipid contents were low when compared with other fish species. All diet assayed produced high liver glycogen accumulation. The recommended protein and lipid levels in diets for red porgy fingerlings were 500 and 150 g kg^?1, respectively.     

Assessment of defined antigens for the diagnosis of bovine tuberculosis in skin test-reactor cattle

The continued use of purified protein derivative (PPD) tuberculin is considered to be the main factor which limits the specificity of diagnostic tests for bovine tuberculosis (TB). This study evaluated a whole blood interferon-gamma (IFN-gamma) assay and compared the diagnostic potential of PPD with two tuberculosis-specific antigens, ESAT-6 and MPB70. To provide estimates of sensitivity and specificity, responses were measured in 180 skin test-reacting cattle, of which 131 were confirmed as tuberculous, and in 128 cattle from TB-free herds. For the skin test reactors, there was a positive correlation between the IFN-gamma responses to PPD from Mycobacterium bovis (PPDB) and PPD from Mycobacterium avium (PPDA), indicating cross-reactivity between these complex antigens which are the basis of the skin test. In comparisons of the ESAT-6 IFN-gamma test with a PPD IFN-gamma test (using PPDB compared with PPDA), there was a decrease in sensitivity (76.3 per cent vs 89.3 per cent), but a clear increase in specificity (99.2 per cent vs 92.2 per cent). The provision of high specificity, even with lower sensitivity, offers major benefits for testing in areas with a low incidence of TB.     

Utility of diagnostic tests used in diagnosis of infection in dogs experimentally inoculated with a North American isolate of Leishmania infantum infantum

Eight female beagles were infected with 1 x 10(7) (low dose, LD) or 2 x 10(8) (high dose, HD) promastigotes of a North American isolate of Leishmania infantum infantum (LIVT-1 strain) isolated from naturally infected Virginia Foxhounds. Two female beagles served as negative controls and 2 male beagles chronically infected (> 3 years) with Leishmania infantum chagasi were positive controls. Bone marrow (BM) and lymph node (LN) aspirates were collected every 6-8 weeks for cytologic evaluation, parasite culture, and polymerase chain reaction (PCR). Serum samples were collected monthly for determination of serologic responses by indirect fluorescent antibody test (IFAT) and diagnostic rK39 antigen. Cultures of BM and LN aspirates and cytology evaluation were consistently positive in positive control dogs during the course of study. Negative control dogs were negative on BM and LN cultures and on cytologic evaluation of aspirates. Amastigotes were present on cytological examination of BM aspirates in 2 experimentally infected dogs. Cultures of LN aspirates were positive on 22 samples, whereas BM cultures were positive on 12 samples for all dogs. IFA titers ranged from 0 to 1 :400 in experimentally infected dogs during the course of the study. Recombinant K39 immunoassay tests were consistently positive in positive control dogs and in the HD dogs by approximately 8 weeks after infection. BM PCR products were identified more consistently in the HD dogs compared with the LD dogs. Kappa statistics indicated PCR correlated better with cultures and cytology than did IFAT or the rK39 immunoassay results in the experimentally infected dogs.     

Successful outcome after surgical correction of large colon atresia in a colt foal

This report describes successful surgical correction and long-term survival of a case of large colon atresia in a 24-h-old Warmblood colt, referred with signs of unrelenting abdominal pain and a suspicion of meconium impaction. Radiographic and ultrasonographic examination was indicative of large colon meconium impaction with secondary ileus. Due to deterioration of the foal, surgery was recommended. An atretic segment was found between the left ventral and dorsal large (ascending) colon. A band of fibrous tissue with no identifiable lumen connected the segments. Surgical correction was done by performing a stapled side-to-side anastomosis. Twelve days after surgery, the foal was discharged. Twenty-two months after discharge, the owner reported the foal developing as expected compared with its peers, but had a mild, self-limiting episode of colic at 20 months of age.